PMID: 7117247Jul 1, 1982Paper

A comprehensive approach to the study of collagen primary structure based on high-performance liquid chromatography

European Journal of Biochemistry
M van der Rest, P P Fietzek

Abstract

A comprehensive approach for the structural microanalysis of collagen based of collagen based on high-performance liquid chromatography (h.p.l.c.) has been developed using calf skin type I collagen as a model system. The alpha, beta and gamma components were separated, after heat denaturation, on a TSK 4000 SW gel permeation column, using a nonvolatile buffer. Monitoring at 210 nm permits the detection of 1 microgram of a single chain. The alpha 1(I) and alpha 2(I) chains were completely resolved using a large-pore reversed-phase column (Vydac 201 TP 4.6) eluted by an aqueous acetonitrile gradient (24-48%) containing 0.01 M heptafluorabutyric acid as an ion-pairing agent. The purified alpha 1(I) chain was digested with CNBr and the resulting fragments separated in the same chromatography system with a gradient containing a 12.8-44.8% acetonitrile gradient. The purified alpha 1(I)CB 3 peptide was further cleaved with trypsin and the resulting peptides separated first by a similar chromatography with a 4-32% acetonitrile gradient. Resolution of some poorly separated peptides was obtained by a rechromatography using trifluoroacetic acid as counterion. The isolated peptides were hydrolyzed and identified by their amino-acid composi...Continue Reading

References

Jan 1, 1976·International Review of Connective Tissue Research·P P Fietzek, K Kühn
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Citations

Sep 1, 1991·International Journal for Parasitology·E A OluochA F Barbet
May 28, 2003·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Z DeylA Eckhardt
Jun 1, 1987·Proceedings of the National Academy of Sciences of the United States of America·D McCormickB R Olsen
May 1, 1994·European Journal of Biochemistry·M Moradi-AméliM Van der Rest
May 16, 2012·Pharmacology & Therapeutics·Diren Beyoğlu, Jeffrey R Idle
Jun 3, 2015·Colloids and Surfaces. B, Biointerfaces·Aleksandra KezwońKamil Wojciechowski
Jul 26, 2012·Biotechnology Journal·Ana Marina FerreiraGianluca Ciardelli
Jan 1, 1988·Annals of the New York Academy of Sciences·M van der RestL Dufour
Jul 1, 1986·American Journal of Medical Genetics·M van der RestF H Glorieux
Mar 17, 1989·Journal of Chromatography·Z Deyl, M Adam
Jun 7, 1996·Biochimica Et Biophysica Acta·J E GerrietsJ A Last
Jan 1, 1985·Journal of Cellular Biochemistry·R MayneW T Butler

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